Cortical hormones



- termediates in the preparation of corticosterone and re- I I Patented Oct. 16, 1956 CORTICAL HORMONES Eugene P. Oliveto, Bloomfield, and Emanuel B. Hershberg, West Orange, N. 1., assignors to Schering Corporation, Bloomfield, N. 1., a corporation of New Jersey No Drawing. Application November 18, 195% Serial No. 469,844

1 Claim. (Cl. 260397.1)

This invention relates to new compounds useful as inlated hormones and to methods of preparing the new compounds. The invention is particularly directed to '3- keto-llfl-hydroxyetiocholanic acid of the formula and its A and A unsaturated analogs.

No instance is recorded in the literature of the preparation of corticosterone via an etio-acid. Fieser and Fieser, in Natural Products Related to Phenanthrene at page 444, say, The diazoketone synthesis cannot beemployed for the synthesis of corticosterone because the llfi-hydroxyl group is eliminated under the conditions required for preparation of the acid chloride and cannot be acylated for protection. However, we have now found conditions which permit the formation of an acid chloride from an etio acid without the simultaneous destruction of an unprotected 1 lfl-hydroxy group.

More particularly, we have found that treatment of metal salts 3-keto-1IB-hydroxyetiocholanic acidand the corresponding A -cholenic and A choladienic acids with oxalyl chloride, preferably at temperatures below about 50 0, results in the production in good yieldtof the corresponding acid chlorides without aflecting the fi-hydroxy group. The reaction may be carried out in an inert organic liquid medium, such as benzene or ether, or the acid salt may be treated directly with the oxalyl chloride.

Making use of this discovery, we have developed a new method for the synthesis of the important adrenal hormone eorticosterone, .a method which is simpler and which proceeds in better yield than previous methods. The reaction scheme is outlined below: XI

' XII The 3,11-diketoetiocholanic acid (IV) can be prepared I in a number of ways:

v I C013 60 (a) Oxidation of pregnan3a-ol-1l,20-'dione (III) with l I alkaline hypohalite solution. O- (b) Ozonization of 21 benzylidenepregnan 3a o1- 11,20-dione (I), which removes the side-chain and oxidizes the 3-hydroxyl to a ketone.

lfldn04 v 65 (c) KMnOI, oxidation of I, which yields 3m-hydroxyll-ketoetiochola'nic acid (H). Mild oxidation of II then 110- HO- gives v I v I r H H V The 3-ketone group of IV must then be protected so 1 v 1 that the ll-ketone can be selectively reduced. This can 70 be done by any number of conventional procedures, e. g. ll l formation of the dimethyl kctal, ethylene glycol ketal,

semicarbazone, oxime, etc.

zones and oximes.

' The 11- ketone is 'then 'r 7 7 group, preferably with sodium or potassium borohydridc.

' Stronger reducing agents (e.-g. lithium borohydrid or lithium aluminum hydride) cannotbe used because they would attack the carboxyl group.

a The protective group at C-3 is then removed by conventional methods, e. g. dilute acid for hydrolysis of the 3- ketals, and pyruvic acid for hydrolysis of the semicarba- The'side-chain is built up by converting the acid group I to an acid chloride (VIII) reacting this'with diazo methane, and then heating the diazoketone (IX) with acetic acid to give the 2l:acetate-20-ketone (X).

The double bond is'introduced into ring A'by the usual methods of bromination and dehydrobromination to give corticosterone acetate XII. Alkaline or acid hydrolysis gives free corticosterone V r There are many variations which may :be used to arriveat the same desired product. For example, the A-ring double bond may be introduced into Compound I IV, the B-ketone protected by either a semicarbazone,

ketal or enol ether, and the ll-ketone reduced with sodium borohydride, viz. V p

educed to the llp-hydroxyl The side-chain'is then built up by theprocedure of the invention: V a

XIX I XII Compound XIX may also be prepared by the introduction of a double bond into Compound VII. Making use of. this new method, it is possible to prepare.

A -pregnadien-11p,21-diol-3,20-dione (XXVII), a new steroid, which possesses even greater cortical hormonal properties than corticosterone. The reaction scheme is outlined below:

xxrv f X V ooomolicw XXVII Example I.--Preparation of 3,11-diketoetiocholkmic acid (IV) A solution of 1.00 g. of 2l-benzyli=denpregnam3wol- 11,20-dione (I) in 150 ml. of C. P. ethyl acetate is ozonized at -70 C. I On the appearance of a permanent bluec'olor the ozone formation is stopped and the system flushed with oxygen ten minutes longer. After treating with 2 g. 'of zinc dust and 5 ml. of acetic acid, the mixture is allowed 'to stand at room temperature overnight, at which/time a starch-iodide test is negative. The.

solids are removed by filtration, washed with ethyl acetate 7 and the filtrate steam distilled to give 0.56 g. of a solid.

The product, crystallized twice from acetone-hexane, melts C OCH OH irared spectrum shows no free hydroxyl group.

Example II.--Preparation of 3,11-diketetiocholanic acid (IV) To a solution of 1.00 g. of I in 35 ml. of 85% aqueous acetone at 40 C. is added instantly a solution of 1.0 g. of potassium permanganate in 60 m1. of 85% aqueous acetone. The mixture is stirred 15 hours at room temperature, filtered, the filtrate heated to its boiling point, and the product ll crystallized by the addition of dilute hydrochloric acid: yield 0.50 g., M. P. 268276 C. d.

Oxidation of II with either chromic acid, N-brom-osuccinimide or N-bromoacetamide gives IV.

Example III.Preparati0n of 3,11-diket0etiocholanic acid 3-ethylene ketal (V) A mixture of 1.00 g. :of IV, ml. of ethylene glycol, 10 ml. of methylene chloride and 1.00 g. of selenium dioxide is stirred overnight at 25 C. A solution of Example I V.Preparati0n of 3-ket0-11B-hydr0xyeti0- cholanic acid (VII) A mixture of 0.60 g. of 3,11-diketoetiocholanic acid 3- ethylene ketal V, 0.20 g. of sodium hydroxide, 30 ml. of C. P. methanol, 0.60 g. of sodium borohydride and 6 ml. of water is refluxed overnight. The methanol is removed under reduced pressure, the clear residue diluted with water, acidified with acetic acid and filtered to give 0.54 g. of 3-keto-1lfl-hydroxyetiocholanic acid 3-ethylene ketal VI, M. P. l60l65 C. This is dissolved in 10 ml. of acetic acid and 10 ml. of water, heated 30 minutes on the steam bath, water added and the mixture chilled and filtered to give 0.44 g. of VII, M. P. 252259 C. Recrystallization from aqueous acetone gives 0.38 g. of VII, M. P. 258261 C., [a] +80.8 C. (ethanol).

Example V.Preparati0n of 3-ket0-11fi-hydr0xy-A etiocholanic acid (XIX) A solution of 3.18 g. of VII in 20 ml. of acetic acid is brominated at room temperature by the dropwise addition of 1.60 g. bromine in ml. of acetic acid. When the bromine color has completely discharged, water is added, and the precipitated 4- bromo-3- keto-llfihydroxyetiocholanic acid is removed by filtration.

A solution of 4.0 g. of the 4-bromide in 40 ml. of acetic acid is treated with 2 g. of semicarbazide base at 0 C. for 4 hours.

The solvent is removed under reduced pressure, water is added, and the 3-semicarbazone of XIX removed by filtration.

The semicarbazone (3.0 g.) is dissolved in 50 ml. of acetic acid and refluxed minutes with an equal weight of pyruvic acid. Removal of the solvent under reduced pressure, followed by addition of water gives XIX, M. P. 250 C.

Example VI.-Preparati0n of 3-keto-11fl-hydroxy- A -eti0ch0ladienic acid (XXIII) A solution of 3.18 g. of VII in 20 ml. of acetic acid is brominated at rom temperature by the dropwise addition of 3.20 g. of bromine in 40 m1. of acetic acid. After the bromine color has been discharged, addition of water precipitates 2,4-dibromo-3-keto-1lfl-hydroxyetiocholanic acid XXII. To 50 ml. of refluxing collidine is added 1.0 g. of the dibromide, and the heating continued one hour.

Chloroform and waterare. added, the organic layer washed several times with..10% sulfuric acid, and then. with water until neutral. The organic layer isdried and evaporated, and the solid residue chromatographed on silica gel to give pure XXIII, M. P. 265 C.

A solution of 603 mg. of sodium hydroxide in 50 ml. of water is used to dissolve 5.02 g. of VII, with the aid of gentle heating. The clear solution is then evaporated to dryness and the residue dried in vacuo for 14 hours to give the sodium salt of VII.

To the salt is added 50 ml. of benzene, the slurry cooled to -10 C., and a solution of 15 ml. of distilled oxalyl chloride in 35 ml. of benzene is added dropwise. and with rapid stirring. The mixture is allowed to warm to room temperature, and stirred 30 minut'es'longer. The solvent is removed under reduced pressure, the residue flushed with 10 ml. of benzene, then dried in vacuo for 30 minutes to give crude 3-keto-1lfl-hydroxyetiocholanyl chloride VIII. This is dissolved in 50ml. of dry methyl: ene chloride and the inorganic residue removed by filtration. The filtrate is cooled to -10? C. and then reacted at this temperature with excess diazomethane (prepared in methylene chloride and dried over sodium hydroxide pellets). The resulting yellow solution is evaporated to dryness in vacuo yielding the light yellow colored diazoketone TX. This is dissolved in 35 ml. of acetic acid and refluxed 10 minutes. Water is added to give a gummy product, which is first dissolved in acetone, the solution taken to dryness and then sludged with ether to give 1.47 g. of X, M. P. -160" C. Two other fractions, 0.28 g., M. P. 148-150 C. and 0.37 g., M. P. 150-152" C., may be obtained giving a total of 2.12 g. This is crystallized from ether to give 1.87 g. of X, M. P. l54157 C.

Example VIII.C0rtic0ster0ne acetate A solution of 1.0 g. of X in 10 ml. of acetic acid is brominated at 20 with 1 molar equivalent of bromine in 5 ml. of acetic acid. The 4-bromide XI is isolated by the addition of water. Without purification this is dehydrohalogenated by the usual semicarbazone procedure to yield corticosterone acetate (XII).

Example IX .-3-ket0-1 1-fi-hydroxy-A -eti0ch0lanic acid XIX Example X.-3-ket0-11B-hydroxy-M-etiocholanic acid The 4-bromide XIII was treated with semicarbazide to give the semicarbazone XIV. This was reduced with sodium borohydride in the usual manner to give XV. Removal of the semicarbazone group with pyruvic acid gives XIX.

Example XI.-C0rtic0sterone acetate Treatment of 1.0 g. of XIX in the form of its sodium salt with oxalyl chloride as described in Example VII gives 3- keto- 11,8-hydroxy- A cholenyl chloride (XX). The chloride is reacted with diazomethane as described in Example VII to give the diazoketone )Qfl which melts with decomposition. Treatment of the diazoketone with acetic and by the method of Example VII gives corticosteronc acetate (XII) in 60% yield.

Example XII.-A -pregnadien-115,21-di0l-3,20-di0ne 21 -acetate XXVI Treatment of 2.0 g. of 3-keto-11B-hydroxy-A -etio' chola'dienicacid XXIII "in the form of its sodium salt with oxalylchloride by the method described in Example VII give 3-keto-41,6 hydroXy-A fi-tiocholadienyl chloride XXIV; The chloride. is ractedwith diazom'etharie as described in Example 'VII to givedhe diazoketoiie' XXV and treatment of the ,diazoketoxieswith acetic acid by the method of Example VII givesXXVI in 55% yie1d,,M. P. 210C. V 7

Example XllI.A ..-pregnadz'en-1 1/5,21.-di0lt .7 3,20-di0fieXXVIL V a A mixture of 1.0 ofXXVLZO mlfof methanol, 0.5 g. of potassium bicarbonate aifid'fi ml. of Water is refluxed 15 minutes under a nitrogen atmosphere. Acetic acid is then added to pH 6; the'metba'iiol is removed under reduced ressure; and water is' added to precipitate A "pregnadieri l'IQZl-dihldoh'(XXVII), M. P.=190 c.

Exam le XlV.V3-ket0-11e hydroryetiocholanyl .;chloride; VIIL. i Y

Amixtui'e'of 1 of the sodium salt of VII (prepared as idExample' VII)" and 15' m1. of distilled oxalyl chlo- 'ducecl pressure and 'n'de is shaken are c. for's hours. During thistime' the steroid gradually goes into solution. The excess solvent' is removed under, reduced pressure and theresiduetdr'ied in vacuo to give crude VII.

Example XV.-'-3'-keto-11,B-hydr0xy-A-cholenyl f chloride XX V A'slu rry of 1 g: of the sodium salt-of XIX ithSOml.

of ether is treated at '25" with 5 ml. o f'distilled oxalyl chloride. The rnixtur'e is allowed to react. at room temperature for 2 hours. The solvent is removed under re the crude acid chloride CX dried in vacuo'. V f

We claim: a 3-keto-1 1B-hydroxy-A etiocholadienic' acid.

References Cited in the file of this patent UNITED STATES PATENTS 2,538,611 Wilde Jan. 16, 19 51 

